RESUMEN
Geigeria poisoning, referred to as 'vermeersiekte' is an important plant poisoning in southern Africa. Three sesquiterpene lactones, isogeigerin acetate (1) ivalin (2) and geigerin (3) were isolated and purified from Geigeria aspera Harv. (Asteraceae). Structures were deduced using 1 and 2D NMR spectroscopy and mass spectrometry, while the absolute configurations of compounds 1 and 3 were determined for the first time by X-ray crystal diffraction analyses. Cytotoxicity of isogeigerin acetate, ivalin and geigerin were compared by exposing a murine skeletal myoblast (C2C12) cell line to varying concentrations of the three sesquiterpene lactones isolated. Cell viability was assessed using the methyl-thiazolyl-tetrazolium (MTT) assay. The EC50s were 3.746, 0.0029 and 3.792 mM for isogeigerin acetate (1), ivalin (2) and geigerin (3), respectively. The results indicate that ivalin is much more toxic, approximately 1000 times, in vitro compared to isogeigerin acetate and geigerin.
Asunto(s)
Geigeria/química , Lactonas/aislamiento & purificación , Lactonas/farmacología , Sesquiterpenos/aislamiento & purificación , Sesquiterpenos/farmacología , Animales , Espectroscopía de Resonancia Magnética con Carbono-13 , Muerte Celular/efectos de los fármacos , Línea Celular , Línea Celular Tumoral , Lactonas/química , Ratones , Espectroscopía de Protones por Resonancia Magnética , Sesquiterpenos/químicaRESUMEN
Vermeersiekte or "vomiting disease" is an economically important disease of ruminants following ingestion of Geigeria (G.) species in South Africa. Sheep are more susceptible, and poisoning is characterized by stiffness, regurgitation, bloat, paresis, and paralysis. Various sesquiterpene lactones have been implicated as the cause of poisoning. The in vitro cytotoxicity of two sesquiterpene lactones, namely, ivalin (purified from Geigeria aspera) and parthenolide (a commercially available sesquiterpene lactone), were compared using mouse skeletal myoblast (C2C12) and rat embryonic cardiac myocyte (H9c2) cell lines, representing the oesophageal, skeletal and cardiac muscles, which are affected in sheep. For 24, 48, and 72 h, both cell lines were exposed. A colorimetric viability assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), was used to assess cytotoxicity. A concentration-dependent cytotoxic response was observed in both cell lines, however, the C2C12 cells were more sensitive, with the half-maximal effective concentrations (EC50s) ranging between 2.7 and 3.3 µM. In addition, the effect that ivalin and parthenolide has on desmin, an important cytoskeletal intermediate filament in myocytes, was evaluated using the C2C12 myoblasts. Disorganization and aggregation of desmin were caused by both sesquiterpene lactones, which could clarify some of the ultrastructural lesions described in vermeersiekte.
Asunto(s)
Citoesqueleto/efectos de los fármacos , Desmina/metabolismo , Lactonas/toxicidad , Mioblastos Esqueléticos/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Sesquiterpenos/toxicidad , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Citoesqueleto/metabolismo , Citoesqueleto/patología , Relación Dosis-Respuesta a Droga , Concentración 50 Inhibidora , Ratones , Mioblastos Esqueléticos/metabolismo , Mioblastos Esqueléticos/patología , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , RatasRESUMEN
Ingestion of large quantities of Geigeria species by sheep causes "vermeersiekte", an economically important poisoning in southern Africa. The toxic principles are several sesquiterpene lactones, such as vermeerin, geigerin and ivalin. These sesquitepene lactones are myotoxic and the disease is characterized by microscopic and ultrastructural lesions in skeletal and cardiac muscle. Murine myoblast cells (C2C12) were exposed to 2.0, 2.5 and 5.0â¯mM geigerin for 24, 48 and 72â¯h to evaluate its effect on cytoskeletal proteins and filaments using immunocytochemistry and immunofluorescence staining. A concentration-dependent cytotoxic response was observed in desmin-expressing murine myoblasts under the light microscope, evidenced by disorganization and dot-like perinuclear aggregation of desmin filaments in the cells. ß-Tubulin, other desmin-associated proteins (αB-crystallin and synemin) as well as the microfilament F-actin were unaffected. The disorganization and aggregation of desmin following exposure to increasing geigerin concentrations is significant and can explain some of the striated muscle lesions observed in "vermeersiekte".